title: Isolation of DNA from oral rinse in HPV positive patients.
abstract: OBJECTIVE
  To extract human genomic DNA from oral rinse in HPV positive patients.
  
  
  METHODS
  Experimental study.
  
  
  METHODS
  Research Laboratory, Ziauddin Medical University, from February to July 2011.
  
  
  METHODS
  Two hundred and fifty oral rinse samples from human papilloma virus (HPV) positive subjects were collected in 50 ml corning tubes. DNA was extracted from 10 ml of oral rinse by using Lysis buffer (sodium dodecyl sulfate, sodium chloride, sodium citrate and EDTA), isopropanol and 3 M sodium acetate and final washing with ethanol. The DNA was quantified by using Qubit dsDNA BR Assay (Qubit 2.0 invitrogen life technologies USA) and the quality was checked by running an aliquot on 0.6% agarose gel stained with ethidium bromide. PCR were done to amplify b-globin gene on chromosome11 by using primers GH20/PCO4 of 260 bp fragment.
  
  
  RESULTS
  The mean concentration of all 250 samples DNA was 15.648 ± 10.50 μg/ml determined by using Qubit 2.0. A single intense band without smearing was seen in almost all cases which confirmed the integrity of DNA. The PCR amplification of human b-globins primers was successfully done.
  
  
  CONCLUSIONS
  The oral rinse method was found a simple and highly appropriate means for non-invasive sample collection with easy storage, DNA recovery and subsequent PCR amplification in HPV positive patients.
authors.affiliation: Department of Biochemistry, Ziauddin Medical University, Clifton, Karachi. haris_lucky@yahoo.com
authors.forename: Muhamamd Haris
authors.initials: MH
authors.lastname: Lucky
authors.forename: Saeeda
authors.initials: S
authors.lastname: Baig
mesh_terms: D004279:DNA, Viral / Q000032:analysis* / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D008297:Male; D009061:Mouth Mucosa / Q000821:virology*; D009067:Mouthwashes; D027383:Papillomaviridae / Q000235:genetics / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000175:diagnosis / Q000821:virology; D016133:Polymerase Chain Reaction; D013048:Specimen Handling; D055544:beta-Globins / Q000032:analysis*
chemical_list: D004279:DNA, Viral; D009067:Mouthwashes; D055544:beta-Globins